Suspension Culture Of MDCK Cells In Bioreactor With Microcarrier And Low Serum Medium

ecause of their high susceptibility to infection with various influenza virus strains, Madin-Darby canine kidney (MDCK) cells have been used as a host for influenza virus isolation and vaccine production. In this work, low serum medium M-LSM containing2～3%serum was designed on base of DMEM/F121:1medium. And this low serum medium was suitable for MDCK cells both in static and microcarrier suspension cultures.MDCK cells were seeded in T-25flasks at the density of0.8×105cells/ml, and the maximum cell density reached17.1×105cells/ml at132h after inoculation. MDCK cells could be directly adapted to low serum medium, and subcultured stably in the medium for more than20passages. Moreover, microcarrier suspension culture was carried out in250ml spinner flask, and during continuous subculture for4times, there was no obvious change in cell growth rate.Furthermore, the culture with medium exchange was performed in1.5L stirred tank bioreactor with3.5g/L microcarriers, and the maximum density of MDCK cells reached37.2×105cells/ml. In another culture, MDCK cells were trypsinzed in digestive bioreactor when the cell density reached19.3×105cells/ml at48h in1.5L bioreactor. Then the cells detached from microcarriers were transferred to5L bioreactor, in which the cell density of36.7×105cells/ml was achieved after culturing for96hours.